The organization of RNA contacts by PTB for regulation of FAS splicing.
The organization of RNA contacts by PTB for regulation of FAS splicing.
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通过PTB组织RNA接触的组织，以调节FAS剪接。
DOI：
10.1093/nar/gku519
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发表时间：
2014-07
影响因子：
14.9
通讯作者：
Smith CW
中科院分区：
文献类型：
--
作者：
Mickleburgh I;Kafasla P;Cherny D;Llorian M;Curry S;Jackson RJ;Smith CW
基因表达的转录后步骤由RNA结合蛋白调节。从高分辨率结构到转录组范围的分析，在表征RNA-蛋白质相互作用方面取得了重大进展。由于固有的技术挑战，较少关注具有多个RNA结合结构域的蛋白质与靶RNA结合的方式。我们已经研究了四个RNA识别基序（RRM）结构域的多聚嘧啶束结合（PTB）蛋白，一个主要的剪接调节剂，与FAS前mRNA的条件下，PTB抑制FAS外显子6剪接。结合拴系羟基自由基探测，有针对性的个别RRM和单分子分析的失活揭示了不平等的分工之间的四个RRM的PTB。RRM 4的RNA结合对于功能是最重要的，尽管固有结合特异性低并且完全缺乏破坏RNA上的单个RRM 4接触点的作用。有序的RRM 3 -4双结构域包装提供了一个延伸的结合表面，用于通过前面接头中的碱性残基在RRM 4处相互作用的RNA。我们的研究结果说明了RRM 4的多个替代低特异性结合构型是如何与阻遏物功能相一致的，只要维持由适当的双结构域包装提供的总体核糖核蛋白结构。
Post-transcriptional steps of gene expression are regulated by RNA binding proteins. Major progress has been made in characterizing RNA-protein interactions, from high resolution structures to transcriptome-wide profiling. Due to the inherent technical challenges, less attention has been paid to the way in which proteins with multiple RNA binding domains engage with target RNAs. We have investigated how the four RNA recognition motif (RRM) domains of Polypyrimidine tract binding (PTB) protein, a major splicing regulator, interact with FAS pre-mRNA under conditions in which PTB represses FAS exon 6 splicing. A combination of tethered hydroxyl radical probing, targeted inactivation of individual RRMs and single molecule analyses revealed an unequal division of labour between the four RRMs of PTB. RNA binding by RRM4 is the most important for function despite the low intrinsic binding specificity and the complete lack of effect of disrupting individual RRM4 contact points on the RNA. The ordered RRM3-4 di-domain packing provides an extended binding surface for RNA interacting at RRM4, via basic residues in the preceding linker. Our results illustrate how multiple alternative low-specificity binding configurations of RRM4 are consistent with repressor function as long as the overall ribonucleoprotein architecture provided by appropriate di-domain packing is maintained.
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影响因子：
12.3
作者：
Cereda M;Pozzoli U;Rot G;Juvan P;Schweitzer A;Clark T;Ule J
通讯作者：
Ule J
影响因子：
14.9
作者：
Gooding C;Edge C;Lorenz M;Coelho MB;Winters M;Kaminski CF;Cherny D;Eperon IC;Smith CW
通讯作者：
Smith CW
影响因子：
4.5
作者：
Amir-Ahmady, B;Boutz, PL;Black, DL
通讯作者：
Black, DL
DOI：
10.1073/pnas.92.4.1113
发表时间：
1995-02-14
影响因子：
11.1
作者：
HEILEK, GM;MARUSAK, R;NOLLER, HF
通讯作者：
NOLLER, HF
影响因子：
11.4
作者：
Cherny, Dmitry;Gooding, Clare;Eperon, Giles E.;Coelho, Miguel B.;Bagshaw, Clive R.;Smith, Christopher W. J.;Eperon, Ian C.
通讯作者：
Eperon, Ian C.